WDR76 degrades RAS and suppresses cancer stem cell activation in colorectal cancer

Eun Ji Ro, Yong Hee Cho, Woo Jeong Jeong, Jong Chan Park, Do Sik Min, Kang-Yell Choi

Research output: Contribution to journalArticle

Abstract

Background: Stabilization of RAS is a key event for the hyper-activation of Wnt/β-catenin signaling and activation of cancer stem cell (CSC) in colorectal cancer (CRC). WD Repeat protein 76 (WDR76) mediates the polyubiquitination-dependent degradation of RAS in hepatocellular carcinoma (HCC). We investigated whether WDR76 destabilizes RAS and acts as a tumor suppressor inhibiting CSC activation in CRC. Methods: We generated mice with deletion of Wdr76 (Wdr76 -/- ) and crosses of Wdr76 -/- with Apc Min/+ (Wdr76 -/- ; Apc Min/+ ) and compared them with wildtype mice (Wdr76 +/+ ) and Apc Min/+ mice (Wdr76 +/+ ; Apc Min/+ ), respectively. Intestinal crypt lengthening, tumorigenesis and CSC activation were analyzed by histology, immunohistochemistry, and immunoblotting. CRC cell line was engineered to stably express or knockdown WDR76 or control vector and was analyzed after spheroid culture. Results: Wdr76 -/- mice, with increased Ras level, displayed crypt elongation and hyper-proliferation. Wdr76 -/- ; Apc Min/+ mice developed more tumors with bigger sizes than Apc Min/+ mice and their tumors showed increased proliferation and CSC activation with elevated RAS and β-catenin levels. In CRC cells, overexpression or knockdown of WDR76 decreased or increased the numbers and sizes of CRC spheroids with inhibition or activation of CSC markers, respectively. In human CRC, lower level of WDR76 was associated with poor patient survival. Conclusions: In analyses of mice with deletion of Wdr76 and CRC spheroids, we found that RAS stability plays important roles in tumorigenesis by affecting proliferation and CSC activation. Our results suggest that destabilization of RAS by WDR76 is a potential strategy for targeting malignant CRC involving CSC activation. Graphic abstract: [Figure not available: See fulltext.]

Original languageEnglish
Article number88
JournalCell Communication and Signaling
Volume17
Issue number1
DOIs
Publication statusPublished - 2019 Jul 30

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Neoplastic Stem Cells
Stem cells
Colorectal Neoplasms
Chemical activation
Proteins
Tumors
Catenins
Carcinogenesis
Neoplasms
Histology
WD40 Repeats
Immunoblotting
Elongation
Hepatocellular Carcinoma
Stabilization
Immunohistochemistry
Cells
Cell Line
Degradation
Survival

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Ro, Eun Ji ; Cho, Yong Hee ; Jeong, Woo Jeong ; Park, Jong Chan ; Min, Do Sik ; Choi, Kang-Yell. / WDR76 degrades RAS and suppresses cancer stem cell activation in colorectal cancer. In: Cell Communication and Signaling. 2019 ; Vol. 17, No. 1.
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abstract = "Background: Stabilization of RAS is a key event for the hyper-activation of Wnt/β-catenin signaling and activation of cancer stem cell (CSC) in colorectal cancer (CRC). WD Repeat protein 76 (WDR76) mediates the polyubiquitination-dependent degradation of RAS in hepatocellular carcinoma (HCC). We investigated whether WDR76 destabilizes RAS and acts as a tumor suppressor inhibiting CSC activation in CRC. Methods: We generated mice with deletion of Wdr76 (Wdr76 -/- ) and crosses of Wdr76 -/- with Apc Min/+ (Wdr76 -/- ; Apc Min/+ ) and compared them with wildtype mice (Wdr76 +/+ ) and Apc Min/+ mice (Wdr76 +/+ ; Apc Min/+ ), respectively. Intestinal crypt lengthening, tumorigenesis and CSC activation were analyzed by histology, immunohistochemistry, and immunoblotting. CRC cell line was engineered to stably express or knockdown WDR76 or control vector and was analyzed after spheroid culture. Results: Wdr76 -/- mice, with increased Ras level, displayed crypt elongation and hyper-proliferation. Wdr76 -/- ; Apc Min/+ mice developed more tumors with bigger sizes than Apc Min/+ mice and their tumors showed increased proliferation and CSC activation with elevated RAS and β-catenin levels. In CRC cells, overexpression or knockdown of WDR76 decreased or increased the numbers and sizes of CRC spheroids with inhibition or activation of CSC markers, respectively. In human CRC, lower level of WDR76 was associated with poor patient survival. Conclusions: In analyses of mice with deletion of Wdr76 and CRC spheroids, we found that RAS stability plays important roles in tumorigenesis by affecting proliferation and CSC activation. Our results suggest that destabilization of RAS by WDR76 is a potential strategy for targeting malignant CRC involving CSC activation. Graphic abstract: [Figure not available: See fulltext.]",
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WDR76 degrades RAS and suppresses cancer stem cell activation in colorectal cancer. / Ro, Eun Ji; Cho, Yong Hee; Jeong, Woo Jeong; Park, Jong Chan; Min, Do Sik; Choi, Kang-Yell.

In: Cell Communication and Signaling, Vol. 17, No. 1, 88, 30.07.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - WDR76 degrades RAS and suppresses cancer stem cell activation in colorectal cancer

AU - Ro, Eun Ji

AU - Cho, Yong Hee

AU - Jeong, Woo Jeong

AU - Park, Jong Chan

AU - Min, Do Sik

AU - Choi, Kang-Yell

PY - 2019/7/30

Y1 - 2019/7/30

N2 - Background: Stabilization of RAS is a key event for the hyper-activation of Wnt/β-catenin signaling and activation of cancer stem cell (CSC) in colorectal cancer (CRC). WD Repeat protein 76 (WDR76) mediates the polyubiquitination-dependent degradation of RAS in hepatocellular carcinoma (HCC). We investigated whether WDR76 destabilizes RAS and acts as a tumor suppressor inhibiting CSC activation in CRC. Methods: We generated mice with deletion of Wdr76 (Wdr76 -/- ) and crosses of Wdr76 -/- with Apc Min/+ (Wdr76 -/- ; Apc Min/+ ) and compared them with wildtype mice (Wdr76 +/+ ) and Apc Min/+ mice (Wdr76 +/+ ; Apc Min/+ ), respectively. Intestinal crypt lengthening, tumorigenesis and CSC activation were analyzed by histology, immunohistochemistry, and immunoblotting. CRC cell line was engineered to stably express or knockdown WDR76 or control vector and was analyzed after spheroid culture. Results: Wdr76 -/- mice, with increased Ras level, displayed crypt elongation and hyper-proliferation. Wdr76 -/- ; Apc Min/+ mice developed more tumors with bigger sizes than Apc Min/+ mice and their tumors showed increased proliferation and CSC activation with elevated RAS and β-catenin levels. In CRC cells, overexpression or knockdown of WDR76 decreased or increased the numbers and sizes of CRC spheroids with inhibition or activation of CSC markers, respectively. In human CRC, lower level of WDR76 was associated with poor patient survival. Conclusions: In analyses of mice with deletion of Wdr76 and CRC spheroids, we found that RAS stability plays important roles in tumorigenesis by affecting proliferation and CSC activation. Our results suggest that destabilization of RAS by WDR76 is a potential strategy for targeting malignant CRC involving CSC activation. Graphic abstract: [Figure not available: See fulltext.]

AB - Background: Stabilization of RAS is a key event for the hyper-activation of Wnt/β-catenin signaling and activation of cancer stem cell (CSC) in colorectal cancer (CRC). WD Repeat protein 76 (WDR76) mediates the polyubiquitination-dependent degradation of RAS in hepatocellular carcinoma (HCC). We investigated whether WDR76 destabilizes RAS and acts as a tumor suppressor inhibiting CSC activation in CRC. Methods: We generated mice with deletion of Wdr76 (Wdr76 -/- ) and crosses of Wdr76 -/- with Apc Min/+ (Wdr76 -/- ; Apc Min/+ ) and compared them with wildtype mice (Wdr76 +/+ ) and Apc Min/+ mice (Wdr76 +/+ ; Apc Min/+ ), respectively. Intestinal crypt lengthening, tumorigenesis and CSC activation were analyzed by histology, immunohistochemistry, and immunoblotting. CRC cell line was engineered to stably express or knockdown WDR76 or control vector and was analyzed after spheroid culture. Results: Wdr76 -/- mice, with increased Ras level, displayed crypt elongation and hyper-proliferation. Wdr76 -/- ; Apc Min/+ mice developed more tumors with bigger sizes than Apc Min/+ mice and their tumors showed increased proliferation and CSC activation with elevated RAS and β-catenin levels. In CRC cells, overexpression or knockdown of WDR76 decreased or increased the numbers and sizes of CRC spheroids with inhibition or activation of CSC markers, respectively. In human CRC, lower level of WDR76 was associated with poor patient survival. Conclusions: In analyses of mice with deletion of Wdr76 and CRC spheroids, we found that RAS stability plays important roles in tumorigenesis by affecting proliferation and CSC activation. Our results suggest that destabilization of RAS by WDR76 is a potential strategy for targeting malignant CRC involving CSC activation. Graphic abstract: [Figure not available: See fulltext.]

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